Assessing the relative stabilities of engineered hemoglobins using electrospray mass spectrometry

被引:23
作者
Apostol, I [1 ]
机构
[1] Baxter Hemoglobin Therapeut, Boulder, CO 80301 USA
关键词
D O I
10.1006/abio.1999.4140
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An ion trap mass spectrometer equipped with an electrospray source was used to examine the relative thermodynamic stabilities of various hemoglobins with respect to both tetramer dissociation and hemin dissociation. The results demonstrated that the stability of hemoglobin molecules can be differentiated by the amount of applied collision-induced dissociation (CID) energy necessary to break up the intact tetramer into its constituent globins. The stability of the intact tetramer was affected by single mutations in the beta-globins. The stabilities of the constituent hologlobins were assessed via trap CID of selected ions. The results demonstrated the importance of the contributions of the hologlobin components to the stability of the intact tetramer. Genetic fusion of two alpha-globins, through the introduction of a single glycine residue between the C-terminus of one alpha-chain and the N-terminus of the second, significantly increased the stability of the hemoglobin pseudo-tetramer. Chemical crosslinking of the beta-globins in addition to genetic fusion of alpha-globins further stabilized the hemoglobin molecule. A dihemoglobin molecule produced by the genetic fusion of two di-alpha-globins with a flexible linker demonstrated a decreased stability relative to the corresponding monohemoglobin. (C) 1999 Academic Press.
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收藏
页码:8 / 18
页数:11
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